This invention relates to compounds having pharmacological activity, processes for their preparation, compositions containing them and methods of treatment 5 involving their use.

In its broadest aspect, the invention is concerned with compounds having the general formula

0 (CH ₂ ) _n -[Y(CH ₂ ) _p ] _q -COR-

in which

R ¹ represents an alkyl or alkenyl C ₁ _ ₁₈ group , _j . optionally substituted by an aryl group or hydroxy or a cycloalkyl C ₃ _ ₆ group,

R ² represents an alkyl C ₁ _ ₁₈ group optionally substituted by an aryl group, or R ² represents a group -CH ₂ -X-R ⁴ wherein X represents 0 or S, and R ⁴ 2 _Q represents hydrogen, an aryl group, or an alkyl or alkenyl ^c J ^rou P optionally substituted by an aryl group,

R ³ represents OH, alkoxy C _1-6 or -NHR ³¹ , wherein R ³¹ represents hydrogen, alkyl C ₁ _ ₆ , OH or ₂ - alkoxy C ₁ _ ₆ optionally substituted by an aryl group.

R ⁵ represents hydrogen, an aryl group or an alkyl or alkenyl C ₁ _ ₁₈ group optionally substituted by an aryl group, n represents 0, 1, 2 or 3, g p represents 0, 1 or 2, q represents 0 or 1,

Y represents -CHR ⁶ -, -CH=CH-, O or S, in which R ⁶ represents hydrogen, an aryl group or an alkyl or alkenyl _l -1 ₈ 9 ^rou P optionally substituted by an aryl group, _T _ and pharmaceutically acceptable derivatives thereof.

According to a first aspect of the invention, there is provided the use of a compound of formula I, as defined above, in the manufacture of a medicament for the treatment or prophylaxis of inflammation. τ_5 According to a second aspect of the invention, there is provided a method of treatment or prophylaxis of inflammation, which method comprises administering a therapeutically effective quantity of a compound of formula I, as defined above, to a patient suffering from or 20 susceptible to an inflammatory condition.

Certain of the compounds of formula I are known for use as pharmaceuticals, or to have pharmacological effects. For example:

European Patent Application No 226304 (Sankyo) 25 describes certain N-acyl a ino acid derivatives which on

concurrent administration with a penem or carbapenem antibiotic reduce the renal toxicity of the antibiotic.

European Patent Application No 136879 (Yamanouchi) describes carboxylic acid and amide derivatives which are 5 orally administered fibrinolytic agents.

Japanese Patent Application No 54-119414 (Fuji) describes the synthesis of _/ S-aminoacid derivatives having antibiotic properties.

German Patent Application No 2252882 (Oreal) relates "L _Q to aminoacid-amine salts containing sulphur which are useful as hair and scalp conditioners.

Yao Hsueh Hsueh Pao 10(7), 418-435 (1963) discloses one compound falling within the scope of formula I which has anthelmintic activity, • _j c A number of compounds active as cholecystographic agents are disclosed in Rec.Trav.Chim.Pays-Bas 87(4) . 308-318 (1968).

In addition there have been a number of publications indicating that certain compounds of formula I may act as 20 inhibitors of various enzymes, but without suggesting any particular utility for those compounds. Examples are J.Med.Chem. 23 , 104-111 (1986), J.Med.Chem. 27, 711-712 (1984), Biochem.Pharmacol. 19(16), 2205-2212 (1980), and Pept. ,Proc.Am.Pept.Symp. ,5th, 209-212 (1977). 25 None of these citations, however, discloses or

suggests any utility of the compounds of formula I as anti-inflammatory agents.

With the above-mentioned exceptions, the pharmaceutical use of the compounds of formula I is novel. 5 As a further aspect of the invention, therefore, there are provided compounds of formula I, as first defined above, and pharmaceutically acceptable derivatives thereof, with the provisos that a) when R ¹ represents methyl, then R ² is other than 0 an alkyl C _1-18 group or benzyl, and b) when R ² represents methyl, then R ¹ is other than an alkyl C _1-18 group, optionally substituted by phenyl, or 2-(5-nitro-2-furyl)ethylenyl, and c) when R ² represents iso-propyl, n represents 1, q τ_5 represents 0 and R ³ represents hydroxy, then R ¹ is other than an alkyl C^g group optionally substituted by . phenyl, and d) when n represents 1, q represents 0, and R ³ represents hydroxy or t-butoxy, then R ¹ and R ² do not

20 both represent CH ₃ (CH ₂ )14-, for use as pharmaceuticals.

In addition to those for which pharmacological effects have previously been described, a number of other compounds of formula I are known. The majority of the compounds are, 25 however, novel and according to a further aspect of the

invention there are provided compounds of formula I, as first defined above, and pharmaceutically acceptable derivatives thereof, with the provisos that i) when R ¹ represents methyl, then R ² is other than an alkyl C _1-18 group or benzyl or 2-phenylethyl or -CH ₂ OH or -CH ₂ SCH ₃ , and ii) when R ² represents methyl, then R ¹ is other than an alkyl C ₁ _ ₁₈ group, optionally substituted by phenyl, or 2-(5-nitro-2-furyl)ethylenyl or 2-(3-phthalimido)propyl or CH ₂ =CH- or CH ₃ CH=CH-, and iii) when R ² represents iso-propyl or ethyl, n represents 1, q represents 0, and R ³ represents hydroxy, then R ¹ is other than an alkyl C^- _ζ group optionally substituted by phenyl, and iv) when R ² represents benzyl, n represents 0, Y represents -CH=CH-, p represents 1, q represents 1, and R ¹ represents 2-methylpropyl, then R ³ is other than hydroxy or methoxy, and v) when R ¹ represents heptadec-8-enyl, n represents 1, q represents 0 and R ³ represents hydroxy, then R ² is other than n-propyl, and vi) when n represents 1, q represents 0, and R ³ represents hydroxy or t-butoxy, then R ¹ and R ² do not both represent CH ₃ (CH ₂ ) ₁₄ -, and vii) when R ¹ represents n-hexyl.

(CH ₂ ) _n [Y(CH ₂ ) _p 3 _q represents (CH ₂ ) ₄ , and R ³ represents hydroxy, then R ₂ is other than n-pentyl.

According to the invention there is also provided a process for the preparation of compounds of formula I and pharmaceutically acceptable derivatives thereof, with provisos i) - vii) above, which process comprises: a) producing a compound of formula I in which R ³ represents hydroxy by hydrolysis of a corresponding compound of formula I in which R ³ represents alkyl C _1→ , or b) producing a compound of formula I in which R ³ represents alkoxy C _1-6 by condensation of an acid of formula II:

R^-COOH II

with an amine of formula III:

NH-CH-(CH ₂ ) _n -[Y(CH ₂ ) _p ] _q - COR- III

wherein R ³ represents alkoxy C _1-6 , or c) producing a compound of formula I in which R ³ represents NHR ³¹ by reacting the corresponding compound

in which R ³ represents COOH with a compound of formula IV

in which R ^JX is as defined above, or d) producing a compound of formula I in which R ³ represents hydroxy or alkoxy C- _j __g by reacting a compound of formula III in which R ³ represents hydroxy or alkoxy ^c l-6 wit ^{11 a} compound of formula V

R ¹ -CO-L V

in which L represents a leaving group, e) producing a compound of formula I in which R ³¹ represents hydroxy by hydrolysis of the corresponding compound in which R ³¹ represents alkoxyC-^.g optionally substituted by an aryl group, or f) producing a compound of formula I in which the chain (CH ₂ ) _n [Y(CH ₂ ) _p ] _q contains a group -CH ₂ CH ₂ - by hydrogenation of the corresponding compound of formula I in which Y represents -CH=CH-, or and where desired or necessary, converting the compound of formula I so obtained to a pharmaceutically acceptable derivative thereof. The hydrolysis of process a) may be carried out under

acid or, preferably, base catalysis. Reagents which may be used for the base catalysed hydrolysis include lithium hydroxide and potassium hydroxide. The reaction may be carried out in the presence of a co-solvent such as tetrahydrofuran.

The reaction of process b) is preferably carried out in an inert solvent such as dichloromethane in the presence of a coupling reagent such as dicyclohexylcarbodiimide with or without 1-hydroxybenzotriazole or other additives. The reaction of process c) is preferably carried out in an inert solvent or mixture of solvents. A coupling reagent such as dicyclohexylcarbodiimide may be employed.

Leaving groups which L may represent in process d) include halide, notably chloride. Acid chlorides of formula V may be prepared by treatment of the corresponding acid with for example, thionyl chloride or oxalyl chloride. The acid chloride is then treated with the amine of formula III in the presence of an inert solvent such as dichloromethane. The reaction of process e) may be carried out in the presence of a suitable catalyst, eg 10% palladium on carbon. Alternatively, the reaction may be carried out by hydrolysis using an inorganic base.

The hydrogenation of process f) may also be carried out over a suitable catalyst, eg 10% palladium on charcoal.

Certain compounds of formula III are novel and useful as intermediates in the preparation of the compounds of formula I. According to another aspect of the invention, therefore, there are provided compounds of formula III

R ^< =

NH-C IH-(CH ₂ ) _n -[Y(CH ₂ ) _p ] _q -COR ₃ ³ III

in which R ³ , R , Y, n, p and q are as first defined above and R ² represents CH ₂ -X-R . A preferred group of such compounds are those in which R represents an aryl group, or an alkyl or alkenyl group optionally substituted by an aryl group. These novel compounds of formula III may be prepared by reacting a compound of formula VI

R ⁴ -X-H VI

with the corresponding compound of formula III in which R ² represents -CH ₂ -L in which L is a leaving group, eg a halogen such as iodine.

Other compounds of formula III, as well as compounds of formulae II, IV, V and VI are either commercially available or may be obtained from commercially available

compounds by known processes which will be apparent to those skilled in the art.

Preferred groups that -^ may represents are alkyl or alkenyl C ₁ -_ ₁₈ , optionally substituted by hydroxy, phenyl or cycloalkyl C ₃ _ ₆ .

Alkyl groups which R ¹ may represent include both straight and branched chain groups. Straight chain alkyl groups include both relatively short chains, eg methyl, ethyl, n-propyl, n-butyl, n-pentyl and n-hexyl, and longer chains, eg pentadecyl, hexadecyl and heptadecyl.

Similarly, branched chains include both relatively simple groups such as i-propyl, i-butyl, s-butyl and t-butyl, and groups containing larger numbers of carbon atoms, eg 6-ethyl-octyl. Alkenyl groups which R ¹ may represent may contain up to 3 double bonds. Where there is more than 1 double bond, they may be conjugated or non-conjugated. Examples of such alkenyl groups include 7-heptadecenyl, 9-pentadecenyl and 2-nonenyl. Cycloalkyl groups with which the group R ¹ may be substituted include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

Aryl groups with which the group R ¹ may be substituted include both carbocyclic and heterocyclic groups. The groups may contain rings of various numbers of

C-atoms and may be fused ring structures. Examples of carbocyclic aryl groups are phenyl and naphthyl. Heteroaryl groups include nitrogen, oxygen or sulphur heterocycles and may contain one or more heteroatoms. Examples of heterocycles containing only one heteroatom include pyrrole, furan, thiophen and pyridine. Groups containing more than one heteroatom include pyrazole, oxazole, thiazole, triazole, oxadiazole, thiadiazole etc. The aryl group may be substituted by a range of substituents including halogen, nitro, alkyl C _x _ ₆ , phenyl and phenyl(alkyl C ₁ _ ₆ ) .

Particularly preferred aryl groups with which R ¹ may be substituted are carbocyclic groups, especially phenyl.

When R ² represents alkyl C^--^, it may be substituted by a similar range of aryl groups to R ¹ . Again, the preferred aryl groups with which R ² may be substituted are carbocyclic groups, especially phenyl.

Preferred groups which R ² may represent include alkyl C _ ₁₈ , more preferably alkyl C ₁ _g, optionally substituted by phenyl, and, more preferably, groups of the formula

-CH ₂ -X-R ⁴ in which X represents S or O, and R ⁴ represents alkyl an aryl group or alkyl C _ 8 substituted by an aryl group.

Alkyl groups which R ^~ ~ may represent include similar groups to those which R ¹ may represent. Similarly, when

R ² represents the group -CH ₂ -X-R ⁴ , R ⁴ may represent a similar range of long and short, straight or branched alkyl groups.

When R ² represents -CH ₂ -X-R ⁴ , then R ⁴ preferably represents an aryl group.

Aryl groups which R ⁴ may represent include carbocyclic groups, notably phenyl, optionally substituted by for example, phenyl and phenylmethyl, and fused ring structures such as naphthyl. Other aryl groups which R may represent include heterocyclic structures. Such heteroaryl groups include nitrogen, oxygen or sulphur heterocycles and may contain one or more heteroatoms. Examples of heterocycles containing only one heteroatom include pyrrole, furan, thiophen and pyridine. Groups containing more than one heteroatom include pyrazole, oxazole, thiazole, triazole, oxadiazole, thiadiazole etc.

The aryl group may be substituted by a range of substituents including halogen, nitro, alkyl C _j .g, phenyl and phenyl(alkyl C^g) .

Alkoxy groups which R ³ may represent include methoxy, ethoxy and propoxy.

When R ³ represents NHR ³¹ , R ³¹ preferably represents hydrogen, alkyl C^ , hydroxy or alkoxy C _1-( -

optionally substituted by aryl. Aryl groups with which R ⁴ may be substituted include both carbocyclic and heterocyclic groups. It is particularly preferred that R ³¹ be substituted by a carbocyclic group, especially phenyl.

Most preferably, R ³¹ represents alkoxy C-^g or, especially, hydroxy.

R ⁵ is preferably lower alkyl, say alkyl C^ , or, more preferably, hydrogen. We prefer compounds in which q is zero.

We also prefer compounds in which n represents 0, 1 o 2, q represents 1 and p represents zero. In such compounds, Y is preferably CHR ⁶ in which R° is preferably hydrogen or alkyl C- _j ^ . We particularly prefer compounds in which n is 1 or 2.

Alkyl groups which R ⁶ may represent include methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl and t-butyl.

Aryl groups with which R ⁶ may be substituted include both carbocyclic and heterocyclic groups. It is particularly preferred that R ⁶ be substituted by a carbocyclic group, especially phenyl.

A particularly preferred group of compounds are those in which R represents alkyl or alkenyl C2_ ₁₈ , optionally

substituted by hydroxy, phenyl or cycloalkyl C ₃ _g,

R ₂ represents alkyl C ₁ _ ₁₈ , optionally substituted by phenyl, or a group -CH ₂ -X-R ⁴ wherein X represents S or 0, and R ⁴ represents alkyl C ₁ _ ₁₈ , an aryl group or alkyl C _ ₁₈ substituted by an aryl group,

R ³ represents hydroxy, alkoxy C^g or NHR ³¹ wherein R ³¹ represents hydrogen, alkyl C-^ , hydroxy or alkoxy C ₁ _g optionally substituted by phenyl, and n represents 0, 1 or 2, q represents 1, and

Y represents -CHR ⁶ -, in which R ⁶ represents hydrogen or alkyl C^g, and pharmaceutically acceptable derivatives thereof. Pharmaceutically acceptable derivatives of the compounds of formula I include esters, amides and salts. Salts of the compounds of formula I include metal ion salts, eg alkali metal and alkaline earth metal salts, and addition salts with suitable bases, eg suitable amines such as dicyclohexylamine and 1-adamantanamine.

The compounds of formula I, and pharmaceutically acceptable derivatives thereof, are useful because they possess pharmacological activity in animals. In particular, the compounds are useful as broad spectrum

anti-inflammatory agents as demonstrated in one or more of the following in vitro assay systems:

1. Porcine pancreatic PLA ₂ induced release of C-araσhidonic acid from mixed micelles: Mixed micelles of l-stearoyl-2-[l- ¹⁴ C]-2-arachidonyl-L-3-phosphatidyl choline and deoxycholate are added to a mixture of porcine pancreatic PLA ₂ and test compound in buffer solution. After 8 minutes the reaction is quenched and ¹⁴ C-arachidonic acid is extracted by the method of Katsumata et al: Anal. Biochem. , 154. 676, (1986), then assayed by scintillation counting techniques.

2. PLA ₂ induced cleavage of a thioester: porcine pancreatic PLA ₂ is incubated with 4-hydroxy-3,5-dioxa- 4,9-dioxo-8-thia-4-phosphahexadecan-l-ammonium hydroxide in buffer solution containing the test compound. Cleavage of the thio ester is assayed photometrically after reaction with DTNB (5,5'-dithiobis[2-nitrobenzoiσ acid]).

3. Human platelet cystolic PLA ₂ induced release of ¹ C-arachidonic acid from liposomes of l-stearoyl-2- ¹⁴ C-arachidonyl phosphatidylcholine in buffer containing 10% DMSO: Assays were carried out as in system 1, but human platelet cystolic PLA ₂ was used in place of porcine pancreatic PLA ₂ .

4. Assay of PLA ₂ in intact HL60 cells and polymorphonuclear leucocytes (PMN) : In the presence of the

test compound and a 5-lipoxygenase inhibitor, HL60 or PMN cells in which the arachidonate moieties of the phospholipids have been equilibrated with tritium-labelled arachidonic acid are challenged with ionophore A23187. After 1 minute (N-formyl-methionyl-leucyl-phenylalanine) or 2 minutes (A23187) the reaction is quenched, the mixture centrifuged and the supernatent assayed by scintillation techniques.

The compounds are indicated for use in the treatment or prophylaxis of inflammatory conditions in mammals including man. Conditions that may be specifically mentioned are: osteoarthritis, rheumatoid arthritis, rheumatoid spondylitis, gouty arthritis and other arthritic conditions, inflamed joints; eczema, psoriasis, dermatitis or other inflammatory skin conditions such as sunburn; inflammatory eye conditions including uveitis and conjunc ivitis; lung disorders in which inflammation is involved, eg asthma, bronchitis, pigeon fancier's disease, farmer's lung, acute respiratory distress syndrome, bacteraemia, endotoxaemia (septic shock) and pancreatitis; conditions of the gastrointestinal tract including aphthous ulcers, gingivitis, Crohn's disease (a condition

of the small and sometimes also of the large intestine) , atrophic gastritis and gastritis varialoforme (conditions of the stomach) , ulcerative colitis (a condition of the large and sometimes of the small intestine) , coeliac disease (a condition of the small intestine) , regional ileitis (a regional inflammatory condition of the terminal ileum) , peptic ulceration (a condition of the stomach and duodenum) and irritable bowel syndrome; pyresis, pain; and other conditions associated with inflammation, particularly those in which phospholipid, lipoxygenase and cyclooxygenase products are a factor.

For the above mentioned uses the doses administered will, of course, vary with compound employed, the mode of administration and the treatment desired. However, in general, satisfactory results are obtained when the compound is administered at a daily dosage of from about O.lmg to about 20mg per kg of animal body weight, preferably given in divided doses 1 to 4 times a day or in sustained release form. For man the total daily dose is in the range of from 7.0mg to l,400mg and unit dosage forms suitable for oral administration comprise from 2.0mg to l,400mg of the compound admixed with a solid or liquid pharmaceutical diluent or carrier.

The compounds of formula I may be used on their own or in the form of appropriate medicinal preparations for

enteral, parenteral or topical administration.

According to the invention there is also provided a pharmaceutical composition comprising preferably less than 80% and more preferably less than 50% by weight of a compound of formula I, or a pharmaceutically acceptable derivative thereof, with the provisos that a) when R ¹ represents methyl, then R ² is other than an alkyl group or benzyl, and b) when R ² represents methyl, then R ¹ is other than an alkyl group, optionally substituted by phenyl, or 2-(5-nitro-2-furyl)ethylenyl, and c) when R ² represents iso-propyl, n represents l, q represents 0 and R ³ represents hydroxy, then R ¹ is other than an alkyl c ^1-6 group optionally substituted by phenyl, and d) when n represents 1, q represents 0, and R ³ represents hydroxy or t-butoxy, then R ¹ and R ² do not both represent CH ₃ (CH ) 4- in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

Examples of such adjuvants, diluents and carriers are for tablets and dragees - lactose, starch, talc, stearic acid; for capsules - tartaric acid or lactose; for injectable solutions - water, alcohols, glycerin,

vegatable oils; for suppositories - natural or hardened oils or waxes.

Compositions in a form suitable for oesophageal administration include tablets, capsules and dragees; compositions in a form suitable for administration to the lung include aerosols, particularly pressurised aerosols; compositions in a form suitable for administration to the skin include creams, eg oil-in-water emulsions or water-in-oil emulsions; compositions in a form suitable for administration to the eye include drops amd ointments.

The compounds of formula I have the advantage that they are less toxic, more efficacious, are longer acting, have a broader range of activity, are more potent, produce fewer side effects, are more easily absorbed or have other useful pharmacological properties, than compounds of similar structure.

The invention is illustrated but in no way limited by the following examples. Example I

5-Methyl-3-r (l-oxohexadecyl)aminolhexanoic acid, dicvclohexylamine salt.

A mixture of 3-amino-5-methylhexanoic acid (0.23g), hexadecanoyl chloride (0.44g) and pyridine (14ml) in

dichloromethane (20ml) was heated under reflux for 4 hours. The solution was poured into water and separated. The organic solution was washed with dilute hydrochloric acid and water, then dried and the solvents removed by rotary evaporation. Purification of the residue by chromatography gave a white solid (0.18g). A solution of the white solid in dichloromethane was treated with dicyclohexylamine to yield the product salt (0.15g) as a white solid after recrystallisation from acetonitrile. M.p.: 100-101.5 ^β C

C ₂₃ H ₄₅ N0 ₃ requires: C 74.41, H 12.13, N 4.96% found: C 74.78, H 11.58, N 4.98%. Example 2 Ethyl 4 ( R)-[ (1-oxohexadecyl.amino]-6-methylheptanoate a) Ethyl 4(R)-rt-butoxycarbonylaminol-6-methylhept-2-enoate To a solution of potassium t-butoxide (1.06g) in dry tetrahydrofuran (30ml) stirred at room temperature under nitrogen was added dropwise over 5 minutes a solution of triethyl phosphonoacetate (2.25ml) in dry tetrahydrofuran (15ml) . After 30 minutes a solution of N-t-butoxycarbonyl- (R)-leucinal in dry tetrahydrofuran (9.48ml of 1M solution) was added and the solution stirred at room temperature for 2 hours then heated under reflux for 3 hours.

The mixture was evaporated, partitioned between dilute hydrochloric acid and ethyl acetate, the organic extract

washed with brine, dried over magnesium sulphate and the solvent removed by rotary evaporation. Chromatography of the residue over silica with 20% ethyl acetate/hexane gave a colourless oil which crystallised on scratching to give the title compound as a white solid (0.41g, 30%). Mass spectrum: m/e = 286 (m+1) . b) Ethyl 4 (R)-r (1-oxohexadecyl)aminol-6-methylhept-2-enoate

A solution of the BOC-protected amine (as prepared above, 1.23mmol, 0.35g) in dichloromethane (20ml) was stirred, cooled to 0°C and treated with trifluoroacetic acid (4ml) . After 19 hours the mixture was evaporated, co-evaporated with toluene and the residue dissolved in ethyl acetate. The solution was washed with saturated aqueous sodium bicarbonate, dried over magnesium sulphate and evaporated to give the crude free amine as a yellow oil. The oil was dissolved in dichloromethane (100ml) , treated with triethylamine (0.172ml) followed by hexadecanoyl chloride (0.338g) and the mixture was stirred for 19 hours. The solution was washed with dilute hydrochloric acid, brine, dried over magnesium sulphate and evaporated. Chromatography of the residue with 20% ethyl acetate/hexane over silica gave the title compound as a white crystalline solid (0.32g, 62%). M.p.: 52-54 ^β C. Mass spectrum: m/e = 423 (M) .

c) Ethyl 4f R)-rf1-oxohexadecyl.amino]-6-methylheptanoate

A solution of the .β-unsaturated ester as prepared in step b) (0.29g) in methanol (50ml) was treated with 10% palladium on charcoal and hydrogenated at atmospheric pressure for 4 hours. The mixture was filtered through hyflo and evaporated to a white solid. Flash chromatography of the solid using ethyl acetate/hexane mixtures afforded the sub-titled compound as a white solid (0.27g, 93%).

M.p.: 55-57°C Mass spectrum: m/e = 425 (M)

Example 3

4fR)-r f1-Oxohexadecyl)amino]-6-methylheptanoic acid

A solution of ethyl 4(R)-[(l-oxohexadecyl)amino]-6- methylheptanoate (0.23g)in tetrahydrofuran (20ml) was mixed with a solution of lithium hydroxide hydrate (0.023g) in water (8ml) and stirred at room temperature for 16 hours.

After evaporation the residue was partitioned between dilute hydrochloric acid and ethyl acetate and the organic layer was separated, dried over magnesium sulphate and evaporated to give the title compound as a white solid

(0.22g, 100%).

M.p.: 67-69'C

Mass spectrum: m/e = 398 (M + 1) , 420 (M + Na)

Example 4 Methyl (E)-5-methyl-3fR)-r fl-oxodec-3-enyl)amino ^~ |hexanoate

A solution of methyl 3(R)-amino-5-methylhexanoate (0.56g) in dichloromethane (10ml) was added over 10 min. to a stirred solution of trans 3-decenoic acid (0.6g) and dicyclohexylcarbodiimide (0.72g) in dichloromethane (20ml) . The mixture was stirred for 16 hours at room temperature when the precipitated solids were removed by filtration. The filtrate was evaporated and the residual oil was purified by flash chromatography on silica to give the title compound as a yellow oil (0.65g). Mass spectrum (Plasma spray) : m/e = 312 (M + H) ⁺ Example 5 fE)-5-Methyl-3(R)-f fl-oxodec-3-enyl)aminolhexanoic acid dicyclohexylamine salt

A solution of the compound from Example 5 (0.53g) in tetrahydrofuran (5ml) was added to a solution of lithium hydroxide monohydrate (O.llg) in water (5ml) and the reaction mixture was stirred for 2 hours at room temperature. The solution was acidified with dilute hydrochloric acid and extracted with ether. The combined ether extracts were washed with water, dried and evaporated. The residual oil was purified by flash chromatography on silica to give a colourless oil (0.35g). A solution of this oil in ethanol (50ml) was treated with dicyclohexylamine to yield the product salt as a white solid (0.4g) .

M. p . : 126-127 ^β C

^C 17 ^H 31 ^N0 3* ^C 12 ^H 23 ^N requires: C 72.75%, H 11.37%, N 5.85% found: C 72.75%, H 11.66%, N 5.87% Mass spectrum: (plasma spray) m/e = 298 (M + H) ⁺ , 182

(M + H) ⁺

The following compounds were prepared by analogous procedures: ii) fZ)-5-methyl-3fR)-r fl-oxooctadec-6-enyl)aminolhexanoic acid dicyclohexylamine salt

M.p.: 88-89 ^β C

^C 25 ^H 47 ^N0 3' ^C 12 ^H 23 ^N requires: C 75.20%, H 11.94%, N 4.74% found: C 75.02%, H 12.06%, N 4.81% ϋi) (Z)-5-methyl-3fR)-[fl-oxohexadec-9-enyl)a ino1hexanoic acid adamantanamine salt

M.p.: 107-108'C

^C 23 ^H 43 ^NO 3- ^C 12 ^H 23 ^N - ⁰ - ^6H 2° requires: C 72.90%, H 11.34%, N 5.15%, H ₂ 0 2.0% found: C 73.14%, H 10.90%, N 5.51%, H ₂ 0 2.1%

Example 6

Ethyl 2.6-dimethyl-4fR)-f f1-oxooctyl)amino]heptanoate a) Ethyl 4fR)-rt-butoxycarbonylamino]-2.6-dimethyl hept-2-enoate A solution of N-t-butoxycarbonyl-(R)-leucinal (2.15g)

in dry dichloromethane (50ml) was stirred under nitrogen at room temperature and treated with a solution of ethyl 2-(triphenylphosphoranylidene)propionate (5.44g) . After 17 hours the mixture was evaporated, treated with hexane-ethyl acetate (10:1, 100ml) and filtered to remove the triphenylphosphine oxide. The filtrate was evaporated and the residue chro atographed over silica with ethyl acetate- hexane (10:1) to give a colourless crystalline solid identified as the sub-titled compound (2.23g). M.p.: 47-48 ^β C

Mass spectrum: (FAB) m/e = 384 (M + Rb) ⁺ α _D = +5.62[C *= 0.53, MeOH] b) Ethyl 2.6-dimethyl-4fR)-r f1-oxooctyl)aminolhept-2-enoate A solution of the product from step a) (1.78g) in dichloromethane (30ml) was stirred at room temperature and treated with trifluoroacetic acid (5ml) . After 19 hours the mixture was evaporated, co-evaporated with toluene and the residue dissolved in ethyl acetate. The solution was washed with saturated aqueous sodium bicarbonate, dried over magnesium sulphate and evaporated to give the crude free amine as a yellow oil. The oil was dissolved in dichloromethane (20ml), treated with triethylamine (0.83ml) followed by octanoyl chloride (1.02ml) and stirred at room temperature for 20 hours. The resulting solution was washed with dilute hydrochloric acid, brine, dried over magnesium

sulphate and evaporated. Chromatography of the residue with 20% ethyl acetate/hexane over silica gave the sub-titled compound as a colourless oil. Mass spectrum: (FAB) m/e = 326 (M + H) ⁺ c) Ethyl 2.6-dimethyl-4fR)-r f1-oxooctyl)amino1heptanoate A solution of the product from step b) (1.3g) was dissolved in methanol (50ml) , treated with 10% palladium on charcoal (0.13g) and hydrogenated at atmospheric pressure for 5 hours. The mixture was filtered through hyflo and evaporated. The mixture of diastereomers was separated by flash chromatography on silica to give the less polar isomer as an oil (0.57g), and the more polar isomer as an oil (0.65g). Mass spectrum less polar isomer: (plasma spray) m/e = 328 (M + H) ⁺

Mass spectrum more polar isomer: (plasma spray) m/e = 328

(M + H) ⁺ Example 7

2.6-Dimethyl-4 R)-T f1-oxooctyl)amino]heptanoic acid

1-adamantanamine salt

A solution of the less polar isomer of the product from Example 6 (0.47g) was dissolved in tetrahydrofuran

(30ml) , stirred at room temperature and treated with a solution of lithium hydroxide hydrate (O.lg) in water

(12ml) . After 16 hours the mixture was evaporated and the

residue partitioned between dilute hydrochloric acid and ethyl acetate and the organic layer separated, washed with brine and dried over magnesium sulphate. The solvent was evaporated and the residue purified by chromatography on silica with 30% ethyl acetate/hexane to give the free acid as a colourless oil (0.38g). The oil was dissolved in dichloromethane (10ml) , treated with 1-adamantanamine (0.177g) and evaporated to give the title salt as a cream foam (0.5g) . Mass spectrum: (FAB) m/e = 451 (M + H) ⁺ C _1? H ₃₃ N0 ₃ .C _1Q H ₁₇ N.0.33H ₂ 0 requires: C 71.04%, H 11.18%, N 6.14%, H ₂ 0 1.27% found: C 71.34%, H 11.25%, N 6.01%, H ₂ 0 1.27%

The more polar diastereomer of the product from Example 6 was treated analogously to give a white foam (0.58g). Mass spectrum: (FAB) m/e = 451 (M + H) ⁺ Elemental analysis: C ₁₇ H ₃₃ NO ₃ .C ₁₀ H ₁₇ N. 0.36H O requires: C 70.94%, H 11.18%, N 6.13%, H ₂ 0 1.40% found: C 70.95%, H 10.96%, N 6.04%, H ₂ 0 1.40% Example 8 Methyl 5-methyl-3 R)-rf1-oxododecyl)amino1hexanoate

A solution of methyl 3(R)-amino-5-methylhexanoate (0.55g) and triethylamine (0.35g) in dichloromethane (20ml) was treated with a solution of dodecanoyl chloride (0.76g)

in dichloromethane (10ml) and the resulting mixture was stirred at room temperature for 16 hours. The organic solution was washed with water, dilute hydrochloric acid, saturated aqueous sodium bicarbonate solution, water, dried and evaporated. The residual oil was purified by flash chromatography on silica to give white crystals (l.lg). M.p.: 49-50'C

Mass spectrum: (electron impact) m/e = 341 M ⁺ Example 9 5-Methyl-3fR)-T 1-oxododecyl)aminolhexanoic acid

A solution of the product from Example 8 (l.lg) in tetrahydrofuran (10ml) was added to a solution of lithium hydroxide monohydrate (0.2g) in water (10ml) and the reaction mixture was stirred for 16 hours at room temperature. The solution was acidified with dilute hydrochloric acid and extracted with ether. The combined ether extracts were washed with water, dried and evaporated. The residual solid was purified by flash chromatography on silica to give the title compound as a white solid. M.p.: 57-58'C

Mass spectrum: (electron impact) m/e = 399 (M - H + TMS) ⁺ C ₁₉ H ₃₇ N0 ₃ requires: C 69.68%, H 11.39%, N 4.28% found: C 69.99%, H 11.20%, N 4.07% The following compounds were prepared by analogous

procedures. ii) 5-Methyl-3(R)-r fl-oxo-10-phenyldecyl)aminolhexanoic acid dicyclohexylamine salt M.p.: 112-113 ^β C C ₂₃ H ₃₇ N0 ₃ .C ₁₂ H ₂₃ N requires: C 75.49%, H 10.86%, N 5.03% found: C 75.55%, H 10.72%, N 4.49% iii) 3fR)-Tf1-Oxohexadecyl)amino]hexanoic acid

M.p.: 96-97 ^β C ^C 22 ^H 4 ₃ ^N0 ₃ ^re< ϊ ^u i ^res: 71.50%, H 11.73%, N 3.79% found: C 71.39%, H 11.52%, N 3.67% iv) fR)-g-f (1-Oxooctyl)aminoIbenzenebutanoic acid M.p.: 100-102 ^P C

C ₁₈ H ₂₇ N0 ₃ requires: C 70.82%, H 8.85%, N 4.59% found: C 70.70%, H 8.94%, N 4.27% v) fR)-β-ffl-Oxohexadecyl)aminoIbenzenebutanoic acid M.p.: 113-114 ^β C

C ₂₆ H ₄₃ N0 ₃ requires: C 74.82%, H 10.31%, N 3.36% found: C 74.75%, H 10.42%, N 3.25% ^v ^- _* 5~Methyl-3(R)-ffl-oxooctyl)aminolhexanoic acid dicyclohexylamine salt M.p.: 134-135*C

^C 15 ^H 29 ^N0 3* ^C 12 ^H 23 ^N requires: C 71.68%, H 11.50%, N 6.19% found: C 71.70%, H 11.48%, N 6.54%

vii) 5-Methyl-3fR)-[ fl-oxo-8-phenyloctyl)aminolhexanoic acid dicyclohexylamine salt

M.p.: 109-110 ^β C

C ₂₁ H ₃₃ N0 ₃ .C ₁₂ H ₂₃ N requires: C 74.95%, H 10.67%, N 5.30% found: C 74.93%, H 10.48%, N 5.27% viii) 5-Methyl-3fR)- f1-oxohexadecyl)aminolhexanoic acid

M.p.: 78-79°C

C ₂₃ H ₄₅ N0 ₃ requires: C 72.01%, H 11.82%, N 3.65% found: C 72.21%, H 11.44%, N 3.56% ix) fE)-5-Methyl-3fR)-r fl-oxodec-4-enyl)aminolhexanoic acid dicyclohexylamine salt

M.p.: 124-125'C

C ₁₇ H ₃₁ N0 ₃ .C ₁₂ H ₂₃ N requires: C72.75: H 11.37; N 5.85% found: C72.41; H 11.25; N 5.81% x) fE)-g R)-r fl-Qxodec-3-enyl)aminoIbenzenebutanoic acid dicyclohexylamine salt

M.p.: 141-142'C

^C 20 ^H 29 ^NO 3* ^C 12 ^H 23 ^N requires: C74.95; H10.22; N4.56% found: C75.05; H9.83; N5.32% xi) fR)-5-Methyl-3rfl-oxo-7-phenylheptyl)aminolhexanoic acid dicyclohexylamine salt M.p.: 200-202"C

^C 20 ^H 31 ^NO 3* ^C 12 ^H 23 ^N requires: C74.66; H10.57; N5.44% found: C74.45, H10.64; N5.07%

Example 10 Methyl 3fR)-f (2-hydroxy-l-oxohexadecyl)amino1-5-methyl- hexanoate

2-Hydroxyhexadecanoic acid (1.3g), followed by

1-hydroxybenzotriazole (0.63g) were added to a stirred solution of methyl 3(R)-amino-5-methylhexanoate (0.74g) in dichloromethane (75ml) . A solution of dicyclohexylcarbodi¬ imide (0.96g) in dichloromethane (25ml) was added and the mixture was stirred for 5 hours. The precipitated solids were removed by filtration and the filtrate evaporated. The residue was taken up in a mixture of diethyl ether and ethyl acetate which was washed with dilute hydrochloric acid, water, saturated aqueous sodium bicarbonate solution, water, dried and evaporated to give a white solid. The mixture of diastereomers was separated by flash chromatography on silica to give the less polar isomer as a white solid. Recrystallisation from hexane gave the title compound as white crystals (0.42g). M.p.: 76-77'C

Mass spectrum: m/e = 414 (M + H) ⁺

The more polar diastereomer was isolated as a white solid (0.55g)

M. p . 87-88 ' C

Mass spectrum: m/e = 414 (M + H) ⁺ Example 11

3fR)-r2-Hydroxy-l-oxohexadecyl) mino!-5-methylhexanoic acid The less polar isomer from Example 10 (0.4g) in tetrahydrofuran (10ml) was treated with a solution of potassium hydroxide (O.llg) in water (5ml). After stirring for 4 hours the tetrahydrofuran was evaporated. The aqueous residue was washed with ether and acidified with dilute hydrochloric acid. The product was extracted with ether which was washed with water, dried and evaporated. Recrystallisation of the residue from ethyl acetate gave the title compound as white crystals (O.llg). M.p.: 76-77'C Mass spectrum: (plasma spray) m/e = 400 (M + H) ⁺ C ₂₃ H ₄₅ N0 ₄ requires: C 69.13%, H 11.35%, N 3.51% found: 68.83%, H 11.33%, N 3.59% The more polar product from Example 10 (0.54g) was treated analogously to give the other diastereomer as white crystals (0.32g) . M.p.: 93'C

Mass spectrum: (plasma spray) m/e = 400 (M + H) ⁺ C ₂₃ H ₄₅ N0 ₄ requires: C 69.13%, H 11.35%, N 3.51% found: C 69.32%, H 11.04%, N 3.60% Example 12

Methyl f4S)-5-T f3-f1.1'-Biphenyl)yl)thiol-4-r f1-oxodec -3-enyl)aminoIpentanoate a) Methyl 4 S)-r f1.1-dimethylethoxy)carbonyl1amino-5- iodopentanoate A solution of methyl (S)-4-[(1,1-dimethylethoxy) carbonyl]-amino-5-hydroxypentanoate (11.8g) [prepared by the method of Shimamoto and Ohfune : Tet Lett 3_0 (29) , 3803-3804 (1989)] in dry tetrahydrofuran (600ml) was treated with N,N'-dicyclohexylcarbodiimide methiodide (31.5g) under nitrogen and heated at 50"C for 5 hours. The mixture was cooled, the solvent evaporated and the residual red oil was taken up in ethyl acetate. The organic solution was washed with saturated aqueous sodium metabisulphite and water then dried (MgS0 ₄ ) and the solvent removed by rotary evaporation. Purification of the residue by chromatography gave a white solid (10.73g) . M.p. 93-95 ^β C Mass spectrum: m/e = 358 (m+H) b) Methyl 5-r3-fl.l'-biphenyl)yl)thio1-4fS)-rr fl.l- dimethylethoxy)carbonyl]amino!pentanoate

A solution of 3-phenylthiophenol (1.58) in dry dimethylformamide (25ml) was stirred under nitrogen and treated, batchwise, with sodium hydride (0.25g of 80% dispersion in oil) at room temperature. After a further -ϊ-hour, a solution of the product of step a) (2.24g) in dry

dimethylformamide (15ml) was added over 1 minute. The mixture was stirred for 3 hours, poured onto dilute hydrochloric acid and extracted with ethyl acetate. The organic solution was washed with water, 50% aqueous sodium metabisulphite solution, water and saturated brine, then dried (MgS0 ₄ ) and the solvent removed by rotary evaporation. Purification of the residue by chromatography gave the sub-title product as an oil (2.29g) which slowly solidified. M.p. 57-59 ^β C

Mass spectrum: m/e = 500 (M+Rb) c) Methyl fE)-5-rf3-fl.l'-biphenyl)yl)thio1-4fS)-rfl- oxodec-3-enyl)amino] entanoate

A solution of the product of part b) , (1.43g) in dichloromethane (50ml) was treated with trifluoracetic acid (12ml) and stirred at room temperature for 18 hours. The excess trifluoracetic acid was evaporated and the residue taken up in dichloromethane. The organic solution was washed with 10% aqueous sodium bicarbonate solution and brine then dried (MgS0 ₄ ) and the solvent evaporated to yield a gum (l.lg).

A solution of the gum (0.55g) in dichloromethane (25ml) was treated with 3-decenoic acid (0.3g), followed by N-hydroxybenzotriazole (0.23g) and then a solution of dicyclohexylcarbodiimide (0.36g) in dichloromethane

(5ml) . The mixture was stirred at room temperature for 18 hours, filtered and the filtrate evaporated. The residue was taken up in ethyl acetate, filtered, and the filtrate washed with dilute hydrochloric acid, water, aqueous sodium bicarbonate solution, water then dried (MgS0 ₄ ) and the solvent removed by rotary evaporation. Purification of the residue by chromatography gave a white solid which was recrystallised from a mixture of hexane and ethyl acetate to give the title product as a white solid (0.13g). M.p.: 61-62 ^* C

Mass spectrum: = m/e = 468 (M+H) C ₂₈ H ₃₇ N0 ₃ S requires: C71.91; H7.97; N3.00; S6.86% found: C71.87; H8.07; N2.67; S6.36% The following compounds were prepared by analogous procedures: ii) Methyl 5-r f3-fl.l'-biphenyl)yl)thiot-4fS)-r fl-oxo-

5-phenylpentyl)amino1pentanoate

M.p.: 96-97'C

C ₂₉ H ₃₃ N0 ₃ S requires: C73.24; H6.99; N2.94; S6.7% found: C73.28; H7.20; N2.71; S6.59% iii) Methyl 5-rf3-fl.l'-biphenyl)yl)thio) 1-4fS)-rf4- cvclohexyl-1-oxobutyl)amino1pentanoate M.p.: 101-102'C C ₂₈ H ₃₇ N0 ₃ S requires: C71.91,; H7.97; N3.00; S6;86% found: C72.05; H8.27; N3.04; S6.48%

iv) Methyl (E)-5-f f2.3,5-6-tetrafluorophenyl)thiol-

4fS)-rfl-oxodec-3-enyl)amino1pentanoate

M.p.: 67-68°C

C ₂₂ H ₂₉ N0 ₃ F ₄ S requires: C57.00; H6.31; N3.02% found: C57.ll; H6.56; N2.94%

Example 13 fE)-5-r f3-f1.1'-Biphenyl)yl)thiol-4 S)-r f1-oxodec

-3-enyl)-aminolpentanoic acid dicyclohexylamine salt

A solution of the compound from Example 12 (0.35g) in tetrahydrofuran (5ml) was added to a solution of lithium hydroxide monohydrate (0.063g) in water (5ml) and the reaction mixture was stirred for 3 hours at room temperature. The tetrahydrofuran was evaporated and the aqueous residue was acidified with dilute hydrochloric acid and extracted with ethyl acetate. The combined organic extracts were washed with water, dried (MgS0 ₄ ) and evaporated.

The residual oil was taken up in dichloromethane

(10ml), treated with dicyclohexylamine (O.llg) and the solvents removed by rotary evaporation. Recrystallisation of the residue from a mixture of ethyl acetate and hexane gave the title product as a white crystalline solid (0.17g)

M.p.: 114-115 ^β C

Mass spectrum: m/e = 454 (M+H) ^C 27 ^H 35 ^N0 3 ^S * ^C 12 ^H 23 ^N

requires: C73.77; H9.21; N4.41; S5.05% found: C73.50; H8.ll; N4.19; S4.99%

The following compounds were prepared by analogous procedures. ii) 5-T3-f1.1'-Biphenyl)yl)thiol-4 S)-rfl-oxo-5-phenyl- pentyl)amino1pentanoic acid. M.p. 94-97'C

C ₂₈ H ₃₁ N0 ₃ S requires: C72.85; H6.77; N3.03; S6.95% found: C73.15; H7.13; N3.12; S7.22% ϋi) 5-rf3-f1.1'-Biphenyl)yl)thiol-4fS)-r f4-cvclohexyl -1-oxobutyl)amino1pentanoic acid. M.p.: 134-135'C

C ₂₇ H ₃₅ N0 ₃ S requires: C71.49; H7.78; N3.09; S7.07% found: C71.09; H.7.99; N3.04; S6.89% iv) 5-r f3- 1.1'-Biphenyl)yl)thiol-4fS)-r fl-oxo-7- phenylheptyl)amino1 entanoic acid M.p.: 95-96.5"C

C ₃₀ H ₃₅ NO ₃ S requires: C73.59; H7.20; N2.86; S6.55% found: C73.30; H7.34; N2.81; S6.70% ^v ) 5-rf3-fl.l'-Biphenyl)yl)thio1-4fS)-rfl-oxo-8- phenyl-octyl)amino]pentanoic acid dicyclohexylamine salt M.p.: 142-143*C C ₃₁ H ₃₇ N0 ₃ S.C ₁₂ H ₂₃ N. 0.65M H ₂ 0 requires: C74.12; H8.87; N4.02; S4.60% found: C74.39; H8.80; N3.93; S4.76%

vi) 5-r f3-f1.1'-Biphenyl)vDthio)-4fS)-r fl-oxo-6- phenylhexyl)-amino]pentanoic acid M.p.: 99.5-100'C

C ₂₉ H ₃₃ N0 ₃ S requires: C73.23; H6.99; N2.94; S6.74% found: C73.48; H6.79; N3.02; S6.95% vii) fE)-4fS)-fl-Oxodec-3-enyl)amino1-5-rT2-f4-Phenyl) thiazolyllthiolpentanoic acid dicyclohexylamine salt M.p.: 141-144"C

^C 24 ^H 32 ^N 2°3 ^S 2* ^C 12 ^H 33 ^N requires: C67.35; H8.64; N6.55% found: C67.23; H8.44; N6.54% viii) E)-5-r f4-fl-Methyl-l-phenyl)ethyl)phenylthiol-4 S)- rfl-oxodec-3-enyl)amino1pentanoic acid

M.p.: 99-102*C C ₃₀ H ₄₁ NO ₃ S requires: C74.51; H9.53; N4.15% found: C74.55; H9.53; N4.11% ix) E)-5-r f2,4-Dif1.1-dimethylethyl)phenyl)thiol-4fS)-

T fl-oxo-dec-3-enyl)amino1pentanoic acid dicyclohexylamine salt M.p.: 121-124'C

^C 23 ^H 46 ^N0 3 ^S * ^C 12 ^H 23 ^N requires: C73.49; H10.38; N4.18% found: C72.83. H10.18; N4.02% x) fE)-5-rf2.3.5.6-Tetrafluorophenyl)thio]-4fS)-rf1- oxodec-3-enyl)amino] entanoic acid dicyclohexylamine salt

M.p. : 130-133 - C

C ₂₁ H ₂₇ N0 ₃ F ₄ S.C ₁₂ H ₂₃ N requires: C62.93; H7.85; N4.45% found: C62.62; H7.85; N4.09% xi) fE)-4fS)-ffl-Oxodec-3-enyl)amino]-5-rf2-thiadiazolyl)- thiol entanoic acid

M.p.: 90-91°C

^C 18 ^H 28 ^N 2°3 ^S 2 requires: C56.22; H7.34; N7.28% found: C56.24; H7.40; N7.29% xϋ) fE)-5-f2-Naphthalenylthio)-4fS)-r fl-oxodec-3-enyl)- amino)pentanoic acid

M.p. 85-87 C

C ₂₅ H ₃₃ N0 ₃ S requires: C70.26; H7.73; N3.28% found: C70.44; H7.76; N2.98% xiii) 5-f2-Naphthalenylthio)-4(S)-ff1-oxododecyl) amino1-pentanoic acid

M.p. 74-76'C

C ₂₇ H ₃₉ N0 ₃ S requires: C70.90; H8.53; N3.06% found: C70.04; H8.43; N3.06% xiv) fE)-5-r 5-Methyl-l.3.4-thiadiazol-2-yl)thiol-4fS)- ri-oxo-dec-3-enyl)amino]pentanoic acid

M.p.: 83-84 ^# C

^C 1 ₈ ^H 2 ₉ ^N ₃ ^S 2 requires: C54.ll; H7.32; NIO.52% found: C54.ll. H7.56; NIO.40% xv) fE)-4fS)-rfl-Oxodec-3-enyl)amino1-5-rf4-pyridinyl)-

thiol entanoic acid M.p.: 146-148 ^β C Mass spectrum m/e = 379 (M+H) xvi) 5-r 3-fl.l'-Biphenyl)yl)thio1-4fS)-rf7-ethyl-l- oxononyl)amino1pentanoic acid M.p. 77-79 ^β C

C ₈ H _3g N0 ₃ S requires: C71.60; H8.37; N2.98; S6.83% found: C71.72; H8.52; N2.87; S6.99% xvii) fE)-4fS)-T l-Qxodec-3-enyl)amino]-5-r f2-pyrimidinyl) thiopentanoic acid M.p.: 96-98 ^β C ^C 19 ^H 29 ^N ₃ ° ₃ ^S requires: C60.08; H7.64; Nil.07% found: C60.16; H7.37; NIO.96% xviii) fE)-5-f 4-f1.1'-Biphenyl)yl)thiol-4fS)-T l-oxodec-3- enyl)amino] entanoic acid M.p.: 120-122 ^β C C ₂₃ H ₃₅ N0 ₃ S requires: C71.52; H7.73; N3.09% found: C71.67; H7.88; N3.13% xix) fE)-4fS)-f fl-Oxo-7-phenylheptyl)amino]-5-r 4-phenyl methyl)phenylthiolpentanoic acid M.p.: 85-88'C C ₃₁ H ₃₇ N0 ₃ S requires: C73.87; H7.35; N2.78% found: C74.10; H7.61; N2.47% xx) fE)-4fS)-r fl-Oxodec-3-enyl)aminol-5-r 5-phenyl-l,2,4- triazol-2-yl)thio]pentanoic acid

M. p . : 139-141 ^β C

C ₂₃ H ₃₂ N ₄ 0 ₃ S requires: C62.13; H7.25; N12.60% found: C61.89; H7.30; N12.33% xxi) fE)-5-T4.5-Diphenyl-2 IH)-imidazolyl)thiol-4 S)- f fl-oxodec-3-enyl)amino]pentanoic acid M.p.: 135-139 ^β C

^C 30 ^H 37 ^N 3°3 ^S - ⁰ - ^67H 2° requires: C67.76; H7.27; N7.90% 2.3% H ₂ 0 found: C68.ll; H7.07; N7.80% 2.3% H ₂ 0 xxii) fE)-5-rr1.1'-Biphenyl)-3-yl1thio]-4 S)-r 1-oxodec-

-4-enyl)amino] entanoic acid

M.p.: 91-93 ^β C

C ₂₇ H ₃₅ N0 ₃ S requires: C71.49; H7.78; N3.09% found: C71.54; H7.71; N3.12% Example 14

Methyl fE)-3fS)-rfl-oxodec-3-enyl)aminol-4- phenylthio)butanoate a) Methyl 3fS)-T f1.1-dimethylethoxy)carbonyl1amino-4- hvdroxybutanoate ^A solution of N-[(1,1-dimethylethoxy)carbonyl]aspartic acid 4-methyl ester (24.6g) in ethyl acetate (400ml) was treated with N-hydroxysuccinimide (11.47g) and cooled to

0 ^β C. A solution of dicyclohexylcarbodiimide (20.57g) in ethyl acetate (250ml) was added slowly and the resulting mixture was stirred at room temperature for 18 hours. The

suspended solids were removed by filtration and the filtrate was evaporated. The residue was dissolved in dry tetrahydrofuran (250ml) and dry ethanol (50ml) , cooled to -5°C and treated batchwise with sodium borohydride (11.31g) over 40 minutes. The mixture was stirred at 0"C for a further 3 hours, quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic extract was washed with brine, dried (MgS0 ₄ ) and the solvent evaporated. Purification of the residue by chromatography gave the sub-title product as a colourless oil (3.6g) Mass spectrum m/e = 318 (M+Rb) b) Methyl 3fS)-f 1.1-dimethylethoxy)carbonyl]amino-4- iodobutanoate The sub-title product was obtained as a yellow solid by treatment of the product of step a) by a procedure analogous to that described in Example 12a) . Mass spectrum m/e = 428 (M+Rb) c) Methyl 3 S)-rr l.1-dimethylethoxy)carbonyl]amino-1- 4-fphenylthio)butanoate

The sub-title product was obtained as a gum from the product of step b) by a procedure analogous to that described in Example 12b) . Mass spectrum m/e = 325 d) Methyl fE)-3fS)-r fl-oxodec-3-enyl)amino]-4-(phenylthio)-

butanoate

The title product was obtained as an oil by treatment of the product of step c) by a procedure analogous to that described in Example 12c) . Mass spectrum m/e = 377

The following compounds were prepared by analogous procedures. ii) Methyl fE)-4-[4-nitrophenyl)thio]-3(S)-[ (l-oxodec-3- eny1)-amino]butanoate M.p.: 72-76'C 21 ^H 30 ^N 2°5 ^S requires: C59.69; H7.16; N6.63% found: C59.90; H7.27; N6.38% iii) Methyl 3fS)-[ fl-oxooctyl)amino]-4-fphenylthio)- butanoate M.p.: 54-44 ^β C 1 ₉ ^H 29 ^N0 3 ^S requires: C64.92; H8.32; N3.98% found: C64.48; H8.13; N3.99% Example 15 fE)-3fS)-r fl-0xodec-3-enyl)amino]-4-f henylthio)- butanoic acid dicyclohexylamine salt

The title product was obtained as a colourless solid by treatment of the product of Example 14d) by the procedure of Example 13a) . M.p.: 127-130'C ^C 20 ^H 29 ^NO 3 ^S ' ^C 12 ^H 33 ^N - ⁰ .17H ₂ O

requires: C70.08; H9.63; N5.11% found: C69.83; H9.55; N5.26%

The following compounds were prepared by analogous procedures: ii) (E)-3fS)-r fl-Oxodec-3-enyl)amino1-4-rf2-dibenzofuran) thiolbutanoic acid dicyclohexylamine salt M.p. 101-103"C

^C 26 ^H 31 ^N0 4 ^S*C 12 ^H 23 ^N requires: C71.89; H8.57; N4.41% found: C71.83; H8.44; N4.38% iii) fE)-4-rf4-Nitrophenyl)thio1-3fS)-rfl-oxodec-3-enyl)- aminolbutanoic acid

M.p.: 113-116 ^β C

C ₂₀ H ₂₈ N O ₅ S requires: C58.80; H6.90; N6.86% found: C58.56; H6.76; N6.60% iv) (E)-3fS)-r fl-Oxodec-3-enyl)amino]-4- henylmethylthio)- butanoic acid dicyclohexylamine salt

M.p. 110-112'C

^C 21 ^H 31 ^N0 3 ^S*C 12 ^H 23 ^N requires: C70.92; H9.74; N5.01% found: C70.81; H9.92; N4.73% v) fE)-4-T fl-Naphthalenylthio1-3fS)-r fl-oxodec-3-enyl)- amino1butanoic acid

M.p.: 119-120'C ^C 24 ^H 31 ^N0 ₃ ^S requires: C69.70; H7.56; N3.39%

found: C69.12; H7.85; N3.26% vi) fE)-4-r f2-f1.1'-Biphenyl)yl)thiol-3fS)-r f1-oxodec-

-3-enyl)amino1butanoic acid

M.p.: 79-81 ^β C ^C 26 ^H ₃ 3 ^N0 ₃ ^S requires: C71.04; H7.57; N3.19% found: C70.62; H7.64; N2.93% vii) 3fS)-fAcetylamino)-4-fhexadecylthio)butanoic acid

M.p.: 91-92 ^β C

C ₂₂ H ₄₃ N0 ₃ S requires: C65.79; H10.79; N3.49% found: C65.84; H10.40; N3/48% viii) 4-r f4-fl.l'-Biphenyl)yl)thiol-3 S)-r fl-oxooctyl)- amino]butanoic acid

M.p.: 156-158'C

C ₂₃ H ₃₁ N0 ₃ S requires: C69.70; H7.55; N3.39% found: C69.60; H7.89; N3.36% ix) 4-ffHexadecyl)thio]-3fS)-f fl-oxooctyl)amino1 butanoic acid

M.p.: 77-78 ^β C

C ₂₈ H ₅₅ N0 ₃ S requires: C69.23; H11.41; N2.88% found: C69.07; H11.89; N2.90% x) fE)-4-rf4-Benzyl)phenylthio) 1-3fS f1-oxodecen-

3-enyl)amino1butanoic acid

M.p.: 71-72'C

C ₂₇ H ₃₅ N0 ₃ S requires: C71.49; H7.78; N3.09% found: C71.27; H7.63; N2.92%

xi) E)-4-r f3-f1.1'-Biphenyl)yl)thiol-3fS)-r 1-oxodec- 3-enyl)amino]butanoic acid M.p.: 94-96 ^β C

C ₂₆ H ₃₃ N0 ₃₃ S requires: C71.04; H7.57; N3.19% found: C70.55; H7.41; N3.08% xii) 3fS)-f l-Oxooctyl)amino1-4-r f2-phenylethyl)thiol- butanoic acid dicyclohexylamine salt M.p.: 120-121'C

^C 20 ^H 31 ^NO 3 ^S * ^C 12 ^H 23 ^H2 requires: C70.28; H9.95; N5.12% found: C70.56; H9.77; N5.20% xiii) 4-rf3-fl.l'-Biphenyl)yl)thio1-3fS)-rfl-oxooctyl)- a ino]butanoic acid

M.p.: 78-80°C ^C 24 ^H ₃ 1 ^N0 ₃ ^S requires: C69.70; H7.55; N3.39% found: C69.48; H7/37; N3.38% xiv) 4-Octylthio-3fS)-rfl-oxooctyl)amino]butanoic acid

M.p.: 71-72'C

C ₂₀ H ₃₉ NO ₃ S requires: C64.34; H10.45; N3.78% found: C64.59; H10.12; N3.93% xv) fE)-4-r2-NaPhthalenylthio1-3fS)-rfl-oxodec-3-enyl)- amino]butanoic acid

M.p.: 70-74 ^β C

C ₂₄ H ₃₁ N0 ₃ S requires: C69.70; H7.55; N3.39; S7.75%

found: C70.03; H7.72; N3.57; S8.01% xvi) 4-r2-Naphtha1enylthio1-3fS)- \ f1-oxooctyl)amino]- butanoic acid

M.p.: 99-100'C ^C 22 ^H 29 ^N0 3 ^S requires: C68.18; H7.54; N3.61% found: C68.53; H7.66; N3.66%

Example 16

Methyl fE)-5-rf3-f1.1'-Biphenyl)vDoxyl-4fS)- f l-oxodec-3-enyl)amino]pentanoate a) Methyl 5-f (3-(l,l'-biphenyl)yl)oxy]-4(S) - T (1,1- di ethylethoxy)carbonyl]amino]pentanoate

A solution of methyl(S)-4-[ (1,1-dimethylethoxy) carbonyl]amino-5-hydroxypentanoate (0.25g) in dry benzene

(10ml) was treated with 3-phenylphenol (0.17g) and triphenylphosphine (0.26g). A solution of diisopropyl azodicarboxylate (0.20g) in dry benzene (10ml) was added over **s-hour. The mixture was stirred for 72 hours and then the solvent was evaporated. The residue was purified by chromatography to give the sub-title product as a colourless oil (0.13g)

Mass spectrum m/e = 400 (M+H) b) Methyl E)-5-r f3-f1,1'-biphenyl)yl)oxyl-4fS)-rf1- oxodec-3-enyl)amino1pentanoic acid

The title product was obtained as a white solid by treatment of the product of step a) by a procedure

analogous to that described in Example 12c) .

M.p.: 65-66'C

C ₂₈ H ₃₇ N0 ₄ requires: C74.47; H8.26; N3.10% found: C74.40; H8.54; N3.17% Example 17 fE)-5-f3-f1.1'-Biphenyl)yl)oxyl-4 S)-r fl-oxodec-3- enyl)amino1 entanoic acid

The title product was obtained as a white solid from methyl (E)-5-[(3-(1,1'-biphenyl)yl)oxy]-4(S)-[ (1-oxodec- 3-enyl)amino]pentanoic acid by a procedure analogous to that described in Example 13a) .

M.p. 88-90'C

C ₂₇ H ₃₆ N0 ₄ requires: C73.94; H8.27; N3.19% found: C73.83; H7.96; N3.25% Example 18 fR)-N-Phenylmethoxy-θ-ff1-oxooctyl)amino- benzenebutanamide

A solution of the compound of Example 9vi) (0.61g,

0.002 moles), benzyl hydroxyla ine hydrochloride (0.32g, 0.002 moles), hydroxybenzotriazole (0.33, 0.002 moles) and triethylamine (0.2g, 0.002 moles) in a mixture of tetrahydrofuran and dichloromethane (20ml, 1:1) was stirred at room temperature for -5-hour. The mixture was cooled to

5 ^β C and treated with dicyclohexylcarbodiimide (0.45g, 0.022 moles) . The resulting solution was allowed to warm to room

temperature, stirred for a further 16 hours and then acetic acid (2 drops) added. The mixture was filtered and the filtrate evaporated to a solid. A solution of the residue in ethyl acetate was washed with dilute hydrochloric acid, water, sodium bicarbonate solution and water. The dried organic solution was evaporated to low volume whereupon crystallisation commenced. The white solid recrystallised from a small volume of methanol to give the product, (650mg, 80%) . M.p.: 152-3'C

^C 25 ^H 34 ^N 2°3 requires: C73.17; H8.29; N6.82% found: C73.17; H8.32; N6.49% Example 19 fR)-N-Hydroxy-fl-ff1-oxooctyl)amino)benzenebutanamide A solution of the compound of Example 18 (400mg) in ethanol (20ml) was stirred over 10% palladium on carbon for 3 hours at room temperature. The solution was filtered and the filtrate evaporated to a solid. The residue was recrystallised from ethyl acetate to give the title product (200mg, 64%) as colourless needles. M.p.: 140-145'C.

^C 1 ₈ ^H 28 ^N 2°3 requires: C67.50; H8.75; N8.75% found: C67.52; H8.87; N8.79% Example 20 fE)-N-T4fS)-rl-Amino-l-oxo-5- 2-naphthalenylthio) ]-

pentyl1dec-3-enamide a) 4fS)-rf1.1-Dimethylethoxy)carbonyl]amino-5-f2- naphthalenylthio)pentanamide

A solution of 4(S)-[ (1,1-dimethylethoxy)carbonyl] amino-5-(2-naphthalenylthio)pentanoic acid (0.5g) in tetrahydrofuran (50ml) was treated with N-hydroxybenzotriazole (0.22g) followed by dicyclohexylcarboddiimide (0.33g). After stirring for 15 minutes ammonia (5ml) was added and the mixture was stirred for a further 20 minutes. The solvents were removed by evaporation and the residue was purified by chromatography to give the sub-titled product as a gum (0.4g) . Mass spectrum m/e = 375 (M+H) b) fE)-N-r4fS)-ri-Amino-l-oxo-5-f2-naphthalenylthio)1- pentyl]dec-3-enamide

The title product was obtained as a white solid by treatment of the product of step a) by a procedure analogous to that described in Example 12c) . M.p. 144-146*C ^C 25 ^H 34 ^N 2°2 ^S requires: C70.42; H7.98; N6.57 found: C69.61; H7.82; N6.38 Example 21 fE)-fR)-N-Hvdroxy-3-rfl-oxodec-3-enyl)amino]benzene butanamide a) fE)-fR)-N-f1.1-Dimethylethoxy)-θ-r fl-oxodec-3-enyl)

-a---m-_-i_--n_--o--_-I _^ b----e---n---z---e_--n---_e-_-b_--u----t---a_---n_--a- --m----i_-_d-_-e-—

A solution of the product of Example 9x) (1.2g), hydroxybenzotriazole (0.48g), triethylamine (0.36g) and 0-(t-butyl)hydroxylamine hydrochloride (0.45g) in dichloromethane (20ml) and tetrahydrofuran (20ml) was stirred at room temperature for h hour. The solution was cooled to 10 ^* C, treated with dicylohexylcarbodiimide (0.74g) and stirred at room temperature with acetic acid (0.1ml), filtered and the filtrated evaporated. The residue was purified by flash chromatography on silica to give the sub-title product as a white solid (l.lg). M.p.: 92-94 ^β C

Mass spectrum (FAB): m/e = 403 (M+H) ⁺ b) fE)-fR)-N-Hydroxy-g-r fl-oxodec-3-enyl)amino1benzene butanamide

A solution of the product (0.83g) of part a) in triflouroacetic acid (40ml) was stirred at room temperature for 16 hours. The solution was made alkaline (pH8) with saturated aqueous sodium bicarbonate solution. The product was extracted with ethyl acetate which was dried (MgS0 ₄ ) and evaporated. Partial purification of the residue by flash chromatography gave a solid which was purified by recrystallisation from ethyl acetate to give the title product as a white solid (0.17g) M.p.: 134-136 ^β C

^C 20 ^H ₃₀ ^N 2°3 requires: C69.33; H8.75; N8.09% found: C69.25; H8.86: N8.28% The following compound was prepared by an analogous procedure: ii) fE)-N-T f4S)-ri-Hvdroxyamino-5-r f3- 1.1'-biphenyl)yl) thio]-1-oxolpentyl1dec-3-enamide M.p.: 102-103°C C ₂₇ H ₃₆ N ₂ 0 ₃ S requires: C69.20; H7.74; N5.98% found: C69.49; H7.66; N5.99% Example 22

Methyl fE)-5-f2-naphthalenylthio)-4fS)-r fl-oxodec-3- enyl)amino]-2-f henylmethyl)pentanoate a) Methyl 5- 2-naphthalenylthio)-4fS)-triphenylmethyl amino)pentanoate A solution of methyl 4(S)-[ (1-dimethylethoxy)carbonyl] amino-5-(2-naphthalenylthio)pentanoate (8.7g) (prepared by a procedure analogous to that described in Example 12b)) in dichloromethane (100ml) was stirred and treated with trifluoroacetic acid (10ml) . After 20 hours the mixture was evaporated. The residue was dissolved in dichloromethane, washed with saturated aqueous sodium bicarbonate, dried over magnesium sulphate and evaporated to give the crude free amine as a light brown oil. The oil was dissolved in dichloromethane (100ml) , treated with triethylamine (4.2ml) followed by triphenylmethyl chloride

(7.0g) and the mixture was stirred for 20 hours. The solution was washed with water, dried over magnesium sulphate and evaporated. Chromatography of the residue with 10% ethyl acetate/hexane over neutral alumina gave the title compound as a colourless gum (8.4g). Mass spectrum: m/e = 532 (M+l) . b) Methyl 5-f2-naphthalenylthio)-2-fphenylmethyl)-4fS) -ftriphenylmethylamino) entanoate

A solution of the product of part a) (2.0g) was dissolved in dry tetrahydrofuran (50ml) and cooled to -78"C under a nitrogen atmosphere. A 1.0M solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran (5.7ml) was added dropwise and stirred for 30 minutes. Benzyl bromide (0.6ml) was added and stirred at -78'C for 5 hours. Aqueous citric acid was added and the mixture extracted with ethyl acetate. The combined extracts were washed with water, dried and evaporated. The residue oil was purified by flash chromatography on neutral alumina to give a colourless gum (1.2g). Mass spectrum: m/e = 622 (M+l) c) Methyl 4fS)-amino-5-f2-naphthalenylthio)-2- fphenylmethyl) pentanoate

A solution of the product of part b) (1.2g) in acetone (50ml) was treated with 2M hydrochloric acid (20ml) . After 5 hours the mixture was evaporated to a small volume. The

residue was partitioned between ether and water and the ethereal layer discarded. The aqueous phase was neutralised with saturated sodium bicarbonate and extracted with ethyl acetate. The combined extracts were dried and evaporated leaving a light brown oil (0.6g). Mass spectrum: m/e = 380 (M+l) . d) Methyl fE)-5-f2-naphthalenylthio)-4 S)-r fl-oxodec-3- enyl)amino1-2- phenylmethyl)-pentanoate

The sub-title product was obtained as an oil by treatment of the product of step c) (0.7g) by a procedure analogous to that described in Example 12c) . The mixture of diastereomers was separated by flash chromatography on silica to give the less polar isomer as an oil (0.21g) . Mass spectrum: (FAB) m/e = 532 (M+l) . The more polar isomer was isolated as an oil (0.25g) Mass spectrum: (FAB) m/e = 532 (M+l) . Example 23 fE)-5- 2-Naphthalenylthio)-4fS)-r fl-oxodec-3-enyl) aminol-2- phenylmethyl)pentanoic acid The title product was obtained as an oil (O.lg) by treatment of the less polar product of Example 23d) (0.26g) by a procedure analogous to that described in Example 11. Mass spectrum (FAB) : m/e = 518 (M+l) ⁺

The more polar product of Example 23d) (0.31g) was treated analogously to give the other diastereomer as an

oil (0. 03g) .

Mass spectrum (FAB): m/e = 518 (M+l) ⁺ . Example 24 fR)-β-TN-Methyl-N-f1-oxooctyl)aminoIbenzenebutanoic acid Prepared by method analogous to Example 11. M.p. 107-9'C C ₁₉ H _2g N0 ₃ requires: C 71.47; H 9.09; N 4.39 % found: C 70.90; H 8.50; N 4.40 %